In order to improve the optimum temperature of lipases, The Penicillum expansum lipase (PEL) gene was mutated by site-directed mutagenesis using overlap extension PCR technique. The recombinant plasmid pPIC3.5K-lip-E83V containing matant gene was expressed in Pichia pastoris GS115. The comparison experiments of the mutant PEL-E83V-GS with the wild-type PEL-GS showed that: the optimum temperature of the mutant (45℃) was higher by 5℃ than that of the wild type. The thermostability of the mutant was similar to the wild type. The enzymatic activity of the mutant was 188 U/mL at 37 ℃, which was 80% that of the wild type at the same conditions. The hydrophobic interaction may be enhanced in the surface region by the hydrophilic amino acid, Glu substituted with the hydrophobic amino acid, Val, and be responsible for the improvement of optimum temperature.

Atherosclerosis ; metabolism ; Cyclooxygenase 2 ; metabolism ; Humans

Objective The objective of this study was to explore the relations of quantitatively measurment of serum alphafetoprotein(AFP)and prognosis in severe viral hepatitis.Methods To retrospectively analyse the different level of quantitatively measurment of serum alphafetoprotein(AFP) and distribution of AFP in 213 patients with severe viral hepatitis.Results The abnormity rate of AFP was 68.1%,the abnormity rate of AFP in acute severe hepatitis patients was significantly less than that in subacute severe hepatitis and chronic severe hepatitis(P＜0.01),the death rate of patients with AFP＜20 μg/L,AFP 20～400 μg/Land AFP＞400 μg/L was 86.8%,58.8% and 20.9% respectively,the survival severe hepatitis parients were 85 cases with AFP(248.0±72.5)μg/L in 213 cases,128 cases with AFP(97.6±50.4)μg/L died(P＜0.01).Conclusion The mortality of severe decreases and the survival rates of severe increases gradually along with the higher level of AFP,and it reveals the higher level of AFP in severe hepatitis has a better prognosis,it is the important reference index of prognostic judgement.

Objective To identify the influencing factors for pediatric nursese' reporting of adverse events at 3A hospitals in Beijing. Methods Adopting the random sampling of ten 3A hospitals with pediatric wards in Beijing and conducting an questionnaire survey of some nurses in those hospitals. Results The possibility of adverse events reporting increased with the awareness of adverse events which was related with their severity. Except for title, educational background, length of service, position and past experiences were not associated with the barriers. The major perceived barrier was fear of colleague relationship brokenup and some other undesirable consequence. Conclusions Better training of awareness of adverse events and changes of the punitive culture by the roots as well as some regulation or policy were the major solutions to improve incident reporting.

Objective To investigate the effect of co-stimulatory molecular CD137 and CD28 on the cell proliferation, IL-2 secretion and cell apoptotic rate of activated T cells in naturally senile mice and subacute senile mice induced by D-galactose. Methods Seven-week-old BALB/c male mice were divided into D-galactose induced subacute senile group (D-gal group) , control group and young group randomly. Subacute senile mice model was established by back hypodermic injection of D-galactose (120 mg/kg, dissolved in 0. 1 ml distilled water) everyday for five month. Control group was established by injection of 0. 1 ml distilled water everyday for five month. Young group was injected with nothing. And 16-month-old BALB/c male mice was taken as aged group. The spleen T cells of each group were isolated and activated in vitro stimulation with ConA + IgG, ConA + CD137mAb or ConA + CD28mAb. The cell proliferation, apoptotic rate and IL-2 concentration in cell culture supernate of T cells were detected. Results (1) The cell proliferation (0.422?0.057, A), IL-2 secretion(0.632?0.066, A)and apoptotic rate(68.0%?2. 4%) of T cells in D-gal group stimulated in vitro with ConA+IgG showed no significant difference when compared with those of aged group. Compared with young and control group, activation of T cell in D-gal and aged groups were significantly decreased; (2) Cell proliferation, IL-2 secretion and cell survival of T cells in D-gal group and aged group were significantly promoted by both ConA + CD137mAb [(0. 639?0. 053, A) , (1.119?0.035,A), (53.3%?2.4)%, respectively] and ConA +CD28mAb. CD137 mAb had less effect on both groups than did CD28 mAb. Conclusions (1) Similar age-associated alterations happen in T cells of both D-gal group and aged group. (2) CD137 and CD28 can promote the activation and survival of T cells in aged and D-gal group. But CD28 has stronger effect on regulation of T cells than CD137.

Objective Some cases of lipid storage myopathy (LSM) are similar to the polymyositis(PM) in clinic.To study the muscle pathology is helpful as to making clear the diagnosis and elucidating the mechanism of the LSM. Methods 14 cases of LSM with high level of CK in serum were analyzed clinically and compared with their muscle morphological and histochemical changes.Results Muscle weakness and fatigue were the main complaints of these patients.Most of them had acute or sub acute onset,some of them had relapse courses.All of the muscle specimens showed typical accumulation of lipid in muscle fibers.Necrotic fibers were found in 9 cases,3 of them showed changes with the courses. Conclusion Muscle biopsies and histochemical studies are necessary to differentiate LMS with PM.The fiber necrosis and CK leakage in LSM may be caused by changes of the inter circumstance and the membrane disturbance of cells.

Objective:To identify the distal and proximal influencing factors of father involvement, and explore the mediating role of father's role perception and the moderating role of depression in the effect of his family-of-origin paternal parenting on father involvement.Methods:Totally 572 fathers with children aged under 12 years old completed the father involvement questionnaire, role of the father questionnaire (ROFQ), parental bonding instrument (PBI) and the center for epidemiologic studies depression scale (CES-D). The Pearson correlation analysis and non-parametric percentile Bootstrap method were conducted via SPSS 21.0.Results:(1)Father involvement (2.86±0.58) was found to be negatively correlated with his own family-of-origin paternal indifference (7.30±3.86) ( r=-0.175, P<0.01), positively correlated with his family-of-origin paternal care (10.46±3.98) ( r=0.291, P<0.01), negatively correlated with his depression (7.07±5.09) ( r=-0.266, P<0.01) and positively correlated with his role perception (61.52±5.92) ( r=0.522, P<0.01). (2)Father's role perception partially mediated the relationship between his family-of-origin paternal care and his own paternal involvement (the mediating effect was 0.015, which was 35.71% of the total effects) and fully mediated the relationship between his family-of-origin paternal indifference and his own paternal involvement (the mediating effect was -0.016). (3)Father's depression played a moderating role between his family-of-origin paternal care and his own role perception ( β=-0.069, P<0.05). And it also played a moderating role between his family-of-origin paternal indifference and his own role perception ( β=0.121, P<0.001). The results of J-N methods showed that the values and significance of simple slopes varied according to father's depression levels in both two models. Conclusion:The paternal parenting that fathers had experienced in his family of origin is an important factor affects his own paternal involvement.In the relationship between father's family-of-origin paternal parenting and his own paternal involvement, father's role perception plays a mediating role and his depression level plays a moderating role.

Objective To explore the influence of plasma homocystine(Hcy) level on the prognosis of acute cerebral infarction(ACI) and the effect of interventional therapy.Methods The plasma Hcy levels of 152 patients with ACI were measured by fluorometric method at

Objective To investigate the status and influential factors of mental health of Pakistani and Bangladeshi peacekeeping forces in Liberia.Methods By random sampling,300peacekeeping officers and soldiers dispatched from Pakistan and Bangladesh(150 each)in Liberia were investigated with Chinese Military Mental Health Scale(CMMHS),Military Mental Maladjustment Scale(MMMS) and Social Support Rating Scale(SSRS),and they were carried out for two times on the 7th day and the 120th day after arrival in peacekeeping mission area.Results The total score and each factor score of CMMHS(except for obsessive-compulsive,anxiety and interpersonal sensitivity)and the total score of MMMS of the peacekeepers at the 7th day after their arrival in mission area were significantly higher than those at the 120th day(P

17α-hydroxylase/17,20-lyase deficiency (17OHD) is a rare cause of congenital adrenal hyperplasia.The patient predominantly presents with low-renin hypertension,hypokalemia,lack of secondary sexual development,and in women with primary amenorrhea,in male with pseudohermaphroditism.We herewith analyse the clinical features of a case of 17OHD diagnosed by gene sequencing.And the etiology,clinical manifestations,genetic features,diagnosis and treatment for 17OHD were reviewed.